Team:BCCS-Bristol/Wetlab/BetaGalactosidaseAssays

From 2010.igem.org

(Difference between revisions)
(Motivation)
(Motivation)
Line 5: Line 5:
In order to properly map the PyeaR part in the range of soil nitrate concentrations that agriColi would be exposed to, we needed to get some data on the sensitivity of the component within that small range of concentrations. To that end, we decided that a β-galactosidase assay would be the most appropriate method of obtaining that data, especially given that Edinburgh were kind enough to provide us with the necessary biobrick to run the tests.
In order to properly map the PyeaR part in the range of soil nitrate concentrations that agriColi would be exposed to, we needed to get some data on the sensitivity of the component within that small range of concentrations. To that end, we decided that a β-galactosidase assay would be the most appropriate method of obtaining that data, especially given that Edinburgh were kind enough to provide us with the necessary biobrick to run the tests.
-
Unfortunately it turned out that the [http://partsregistry.org/Part:BBa_K216009 BBa_K216009] part was not entirely appropriate for the assay - the data could be limited by the cellular expression of the LacZΩ portion of β-galactosidase which is under the regulation of a separate promoter on the chromosomal DNA. So we decided to build our own biobrick to enable us to run the tests (see the page on design of the [[:BCCS-Bristol/Wetlab/Part_Design/BioBricks/PyeaR%2BLacZ|PyeaR+LacZ construct]] for more details )
+
Unfortunately it turned out that the [http://partsregistry.org/Part:BBa_K216009 BBa_K216009] part was not entirely appropriate for the assay - the data could be limited by the cellular expression of the LacZΩ portion of β-galactosidase which is under the regulation of a separate promoter on the chromosomal DNA. So we decided to build our own biobrick to enable us to run the tests (see the page on design of the [[:Team:BCCS-Bristol/Wetlab/Part_Design/BioBricks/PyeaR%2BLacZ|PyeaR+LacZ construct]] for more details )

Revision as of 10:05, 24 October 2010

β-Galactosidase Assays

Motivation

In order to properly map the PyeaR part in the range of soil nitrate concentrations that agriColi would be exposed to, we needed to get some data on the sensitivity of the component within that small range of concentrations. To that end, we decided that a β-galactosidase assay would be the most appropriate method of obtaining that data, especially given that Edinburgh were kind enough to provide us with the necessary biobrick to run the tests.

Unfortunately it turned out that the BBa_K216009 part was not entirely appropriate for the assay - the data could be limited by the cellular expression of the LacZΩ portion of β-galactosidase which is under the regulation of a separate promoter on the chromosomal DNA. So we decided to build our own biobrick to enable us to run the tests (see the page on design of the PyeaR+LacZ construct for more details )